PMID- 22537819
OWN - NLM
STAT- MEDLINE
DCOM- 20120910
LR  - 20191210
IS  - 1872-8359 (Electronic)
IS  - 0167-7012 (Linking)
VI  - 90
IP  - 1
DP  - 2012 Jul
TI  - Rapid detection of viable Bacillus pumilus SAFR-032 encapsulated spores using 
      novel propidium monoazide-linked fluorescence in situ hybridization.
PG  - 15-9
LID - 10.1016/j.mimet.2012.04.006 [doi]
AB  - The survival of Bacillus pumilus SAFR-032 spores to standard industrial clean 
      room sterilization practices necessitates the development of rapid molecular 
      diagnostic tool(s) for detection and enumeration of viable bacterial spores in 
      industrial clean room environments. This is of importance to maintaining the 
      sterility of clean room processing products. This paper describes the effect of 
      propidium monoazide (PMA) on fluorescence in situ hybridization (FISH) for 
      detecting and enumerating B. pumilus SAFR-032 viable spores having been 
      artificially encapsulated within poly(methylmethacrylate) (Lucite, Plexiglas) and 
      released via an organic solvent (PolyGone-500). The results of the PMA-FISH 
      experiments discussed herein indicate that PMA was able to permeate only the 
      compromised coat layers of non-viable spores, identifying PMA treatment of 
      bacterial spores prior to FISH analysis as a novel method for selecting out the 
      fraction of the spore population that is non-viable from fluorescence detection. 
      The ability of novel PMA-FISH to selectively distinguish and enumerate only the 
      living spores present in a sample is of potential significance for development of 
      improved strategies to minimize spore-specific microbial burden in a given 
      environment.
CI  - Copyright (c) 2012 Elsevier B.V. All rights reserved.
FAU - Mohapatra, Bidyut R
AU  - Mohapatra BR
AD  - Biotechnology and Planetary Protection Group, Jet Propulsion Laboratory, 
      California Institute of Technology, Pasadena, CA 91109, USA. bmohapatra@disl.org
FAU - La Duc, Myron T
AU  - La Duc MT
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20120417
PL  - Netherlands
TA  - J Microbiol Methods
JT  - Journal of microbiological methods
JID - 8306883
RN  - 0 (Azides)
RN  - 0 (propidium monoazide)
RN  - 36015-30-2 (Propidium)
SB  - IM
MH  - Azides/chemistry
MH  - Bacillus/chemistry/*cytology
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - *Microbial Viability
MH  - Propidium/analogs & derivatives/chemistry
MH  - Spores, Bacterial/chemistry/*cytology
MH  - Staining and Labeling
EDAT- 2012/04/28 06:00
MHDA- 2012/09/11 06:00
CRDT- 2012/04/28 06:00
PHST- 2012/02/02 00:00 [received]
PHST- 2012/04/08 00:00 [revised]
PHST- 2012/04/09 00:00 [accepted]
PHST- 2012/04/28 06:00 [entrez]
PHST- 2012/04/28 06:00 [pubmed]
PHST- 2012/09/11 06:00 [medline]
AID - S0167-7012(12)00140-6 [pii]
AID - 10.1016/j.mimet.2012.04.006 [doi]
PST - ppublish
SO  - J Microbiol Methods. 2012 Jul;90(1):15-9. doi: 10.1016/j.mimet.2012.04.006. Epub 
      2012 Apr 17.